36 STAINING METHODS 



This gives a very sharp differentiation of bacteria and nuclear structures. It 

 has a purplish tinge. Fixation by heat gives the best staining. Allow the stain to 

 act for two to ten minutes. It should not be used until after standing twenty-four 

 hours, and after standing about two weeks it appears to lose its sharp staining power. 



Archibald's Stain. This is an excellent bacterial stain and has 

 been highly recommended by Blue and McCoy in plague work. 



Solution No. i. Solution No. 2. 



Thionin, 0.5 Methylene blue, 0.5 



Phenol crys., 2.5 Phenol cry s., 2.5 



Formalin, i.o Formalin, i.o 



Water, 100.0 Water, 100.0 



Dissolve for twenty-four hours. Mix equal parts and filter. Stain smears 

 fixed by heat or otherwise for ten seconds. 



Nicolle's Carbol Thionin. 



Sat. sol. thionin in 50% alcohol, 10 c.c. 



Carbolic acid solution (2%), 100 c.c. 



Pappenheim's Stain. Take a very small portion of methylene green on the point 

 of a penknife and shake it into a test-tube. Then take up twice as much pyronin 

 and deposit it in the same test-tube. Fill the test-tube one-half full with water and 

 the solution should have a distinct reddish-violet color. A drop on a piece of filter 

 paper shows a violet center and peripheral green ring. The solution should be fresh. 

 Stain from two to five minuted. Differentiate with a little resorcin on a penknife 

 point dissolved in one-quarter of a test-tube full of alcohol. Dehydrate, clear 

 and mount. Polymorphonuclear nuclei stain greenish; nuclei of mononuclears and 

 plasma cells from bluish-red to dull violet. Cytoplasm of lymphocytes and plasma 

 cells purplish-red. Bacteria red. 



Romanowsky Stains. See under section on Blood. For mount- 

 ing specimens showing chromatin staining, as malarial parasites, try 

 panosomes, intestinal flagellates etc., liquid petrolatum is to be highly 

 recommended. The chromatin staining lasts without any fading for 

 at least two years. The acidity of balsam causes rapid fading of the 

 chromatin. 



Neisser's Stain for Diphtheria Bacilli. 



Solution No. i. Solution No. 2. 



Methylene blue, o.igram. Bismark brown, 0.2 



Alcohol, 2 c.c. Water (boiling), 100 c.c. 



Glacial acetic acid, 5 c.c. Dissolve the stain in the boiling 



Distilled water, 95 c.c. water and filter. 



Dissolve the methylene blue in the 

 alcohol and add it to the acetic acid 

 water mixture. Filter. 



